PROJECT SUMMARY: Pancreatic cancer accounts for approximately 3% of all cancers in the United States and approximately 7% of all cancer related deaths. New treatment paradigms are direly needed. Among the emerging treatment strategies, advances in tumor ablation techniques have shown significant promise in both clinical and pre-clinical cancer studies. This proposal will focus on histotripsy, which is the first completely non-invasive, non-thermal, and non-ionizing ablation method capable of overcoming many of the limitations of the majority of other tumor ablation strategies. Histotripsy is an ultrasound ablation method that destroys tissue through the precise control of acoustic cavitation that can selectively destroy tumor tissue while preserving critical structures within the target organ. This method has shown significant initial promise, but has never been evaluated in the pancreas. To date, the lack of clinically and physiologically relevant pre-clinical pancreatic cancer models has been a significant limitation to the development of therapeutic strategies, including histotripsy. To circumvent this limitation, our research team has developed a novel immunodeficient pig model (RAG2/IL2RG) that is ideal for human tumor xenograft studies that we will adapt to study pancreatic cancer in situ. The objective of this proposal is to develop histotripsy as a non-invasive, non-thermal, and image-guided ablation method for in situ ablation of pancreatic tumors. Here, we will implant human pancreatic tumor cells (Panc01) into specific regions of the pancreas of our unique immunodeficient pigs to generate pre-clinical animal models of pancreatic cancer that are superior to any of the current state-of-the-art animal models available to study this disease. We will then use these animals to refine and optimize our histotripsy treatment strategies. Our overarching hypothesis is that histotripsy can achieve safe and selective ablation of pancreatic tumors without inducing unwnanted clinical side effects. Specific Aim 1 will develop a histotripsy treatment strategy for achieving highly targeted pancreas ablation. This Aim will test the hypothesis that a histotripsy treatment strategy can be developed to achieve safe and selective ablation of pancreatic tissue. The long-term safety of histotripsy ablation of pancreas tissue near critical structures will also be evaluated in this Aim. Specific Aim 2 will utilize histotripsy to ablate tumors in an orthotopic porcine model of pancreatic cancer. This Aim will test the hypothesis that histotripsy is capable of selectively imaging and treating orthotopic pancreatic tumors in our unique porcine models. Here, we believe that the orthotopic pancreatic tumor will demonstrate significantly altered mechanical properties and unique changes in the tumor microenvironment compared to the healthy tissue that can significantly impact treatment strategies, which are best evaluated in situ in a large mammal model. This work is significant as it could shift treatment paradigms associated with pancreatic cancer and provide the research community with a new model for therapeutic strategy evaluation.